Human neutralizing monoclonal antibody against rsv f protein and application thereof

Abstract

The application discloses a full human neutralizing monoclonal antibody against RSVF protein and application thereof, and belongs to the field of virology and immunology detection. The monoclonal antibody C16 is screened by using flow sorting-single cell PCR technology, the monoclonal antibody can effectively neutralize RSV infection Hep2 cells (IC 50 =153.10 ng / mL), can be applied to an antibody drug for treating RSV, can be used for developing a diagnostic kit for RSV in IFA, ELISA and colloidal gold chromatography test strip, and can effectively promote and apply RSV differential diagnosis. The application has great industrial production potential.

Description

Technical Field

[0001] This invention relates to a fully human neutralizing monoclonal antibody against RSVF protein and its applications, belonging to the fields of virology and immunology detection. Background Technology

[0002] Respiratory syncytial virus (RSV) is an enveloped, single-stranded, negative-sense RNA virus that readily forms cellular syncytia, making it highly contagious. It is primarily transmitted through droplets and contact, with peak incidence in winter and spring. Typical initial symptoms resemble a cold, including runny nose, cough, and low-grade fever. Infants and young children are prone to developing bronchiolitis and pneumonia, exhibiting severe symptoms such as wheezing, rapid breathing, and feeding difficulties. The elderly and those with underlying medical conditions are more susceptible to pneumonia and respiratory failure. Its dangers extend beyond the risk of acute, severe illness and death; it can also increase the long-term risk of recurrent wheezing and asthma in children. Current treatment focuses on symptomatic and supportive care, with severe cases requiring respiratory support. Prevention includes daily protection, environmental disinfection, and the use of long-acting monoclonal antibodies for high-risk infants. Vaccination is recommended for the elderly and pregnant women, significantly reducing the risk of severe illness and hospitalization.

[0003] RSV has seven main structural proteins (F protein, G protein, SH protein, M protein, N protein, P protein, and L protein). Among them, the F protein is responsible for membrane fusion, and the G protein mediates viral adhesion, making them the main targets for vaccine and antibody development. This invention aims to design a specific binding probe using the RSV F protein and obtain monoclonal antibodies with high affinity and excellent neutralizing activity from the peripheral blood of recovered RSV patients using single-cell PCR technology. The goal is to provide a fully human monoclonal therapeutic antibody with good protective efficacy against RSV. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a fully human monoclonal antibody C16 against RSV F protein and its applications. Because the C16 monoclonal antibody exhibits good specificity for RSV, it can be used to establish immunodiagnostic techniques for RSV, such as ELISA detection methods or to prepare rapid detection methods using colloidal gold immunochromatographic test strips.

[0005] This invention is achieved through the following technical solution: The first objective of this invention is to provide a fully human neutralizing monoclonal antibody against RSVF protein, wherein the monoclonal antibody is composed of a heavy chain and a light chain, wherein the amino acid sequences of the three CDR regions of the variable region of the heavy chain, namely CDR1, CDR2 and CDR3, are as shown in SEQ ID NO. 1, 2 and 3, respectively; and the amino acid sequences of the three CDR regions of the variable region of the light chain, namely CDR1, are as shown in SEQ ID NO. 4, CDR2, and CDR3, are as shown in SEQ ID NO. 5.

[0006] In one embodiment of the present invention, the amino acid sequences of the four FR regions of the heavy chain in the monoclonal antibody, namely FR1, FR2, FR3, and FR4, are shown in SEQ ID NO. 6, 7, 8, and 9, respectively; and the amino acid sequences of the four FR regions of the light chain, namely FR1, FR2, FR3, and FR4, are shown in SEQ ID NO. 10, 11, 12, and 13, respectively.

[0007] In one embodiment of the present invention, the monoclonal antibody has a heavy chain having an amino acid sequence as shown in SEQ ID NO.14 and a light chain having an amino acid sequence as shown in SEQ ID NO.15.

[0008] A second object of the present invention is to provide a nucleic acid molecule encoding the monoclonal antibody.

[0009] In one embodiment of the invention, the nucleic acid molecule encoding the heavy chain in the monoclonal antibody has: (a) a nucleotide sequence as shown in SEQ ID NO. 16; or, (b) a complementary nucleotide sequence to the nucleotide sequence shown in SEQ ID NO. 16; or, a nucleotide sequence that encodes the same protein as (a) or (b) but is different from the nucleotide sequence of (a) or (b) due to the degeneracy of the genetic code.

[0010] In one embodiment of the invention, the nucleic acid molecule encoding the light chain in the monoclonal antibody has: (c) a nucleotide sequence as shown in SEQ ID NO. 17; or, (d) a complementary nucleotide sequence to the nucleotide sequence shown in SEQ ID NO. 17; or, a nucleotide sequence that encodes the same protein as the nucleotide sequence of (c) or (d), but is different from the nucleotide sequence of (c) or (d) due to the degeneracy of the genetic code.

[0011] A third objective of the present invention is to provide a vector for expressing the monoclonal antibody.

[0012] A fourth object of the present invention is to provide a cell expressing the monoclonal antibody.

[0013] A fifth object of the present invention is to provide a medicament against respiratory syncytial virus, the medicament comprising the monoclonal antibody and pharmaceutically acceptable excipients.

[0014] A sixth object of the present invention is to provide a diagnostic kit for diagnosing respiratory syncytial virus, the kit comprising the monoclonal antibody.

[0015] The beneficial effects of this invention are: This invention utilizes flow cytometry-single-cell PCR technology to screen and obtain the fully human monoclonal antibody C16. This monoclonal antibody can effectively neutralize RSV infection of Hep2 cells (ICV). 50 (153.10 ng / mL), it can be used as an antibody drug for the treatment of RSV, and can also be used in IFA, ELISA, and colloidal gold chromatography test strips to develop diagnostic kits for RSV, which can effectively promote and apply RSV differential diagnosis. It has great potential for industrial production. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0017] Figure 1 Memory B cells that are specifically bound by flow cytometry.

[0018] Figure 2 This is an agarose gel electrophoresis pattern of antibody light and heavy chain genes.

[0019] Figure 3 The output image shows the sequence search results for the C16 antibody variable region.

[0020] Figure 4 This is a molecular sieve chromatography purification diagram of the C16 antibody.

[0021] Figure 5 This is the SDS-PAGE pattern of the C16 antibody.

[0022] Figure 6 This refers to the neutralizing ability of the C16 antibody against RSV virus. Detailed Implementation

[0023] The present invention will be further illustrated below with specific examples. These embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, after reading the teachings of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0024] the term: Unless otherwise defined, all technical terms used herein have the same meaning as understood by one of ordinary skill in the art. For definitions and terminology in this field, those skilled in the art may refer to Current Protocols in Molecular Biology (Ausubel). The abbreviations for amino acid residues are the standard 3-letter and / or 1-letter codes used in this field to refer to one of the 20 commonly used L-amino acids.

[0025] In this application, the term "respiratory syncytial virus" or "RSV" refers to the genus Pneumovirus of the family Paramyxoviridae. This virus is a common viral pathogen that causes acute lower respiratory tract infection (ALRI) in infants, the elderly, and immunocompromised adults, and can cause interstitial pneumonia and bronchiolitis.

[0026] In this application, the term "antibody" refers to a protein composed of one or more polypeptides that specifically bind to antigens. One form of antibody constitutes the basic structural unit of an antibody. This form is a tetramer, which consists of two pairs of identical antibody chains, each pair having a light chain and a heavy chain. In each pair of antibody chains, the variable regions of the light and heavy chains work together to bind the antigen, while the constant regions are responsible for the antibody's effector function.

[0027] The "variable region" of an antibody heavy or light chain is the N-terminal maturation region of that chain. Currently known antibody types include κ and λ light chains, as well as α, γ (IgG1, IgG2, IgG3, IgG4), δ, ε, and μ heavy chains, or their other equivalent types. A full-length immunoglobulin "light chain" (approximately 25 kDa or approximately 214 amino acids) contains a single molecule composed of NH2... - A variable region consisting of approximately 110 amino acids at the end, and a COOH group. - The κ or λ constant region at the end. The full-length immunoglobulin “heavy chain” (approximately 50 kDa or approximately 446 amino acids) also contains a variable region (approximately 116 amino acids) and one of the heavy chain constant regions, such as γ (approximately 330 amino acids).

[0028] In this application, the term "antibody" includes any isotype of antibody or immunoglobulin, or antibody fragment that maintains specific binding to an antigen, including but not limited to Fab, Fv, scFv, and Fd fragments, chimeric antibodies, humanized antibodies, single-chain antibodies, and fusion proteins comprising the antigen-binding portion of an antibody and non-antibody proteins. Antibodies can be labeled and detected, for example, by means of radioisotopes, enzymes that produce detectable substances, fluorescent proteins, biotin, etc. Antibodies can also be bound to solid-phase carriers, including but not limited to polystyrene plates or beads. In this application, the term "human monoclonal antibody" refers to an antibody that contains a CDR region derived from a non-human antibody, and other portions of the antibody molecule are derived from one (or more) human antibodies. Furthermore, to preserve binding affinity, some residues in the backbone (called FR) segment may be modified.

[0029] In this application, the term "monoclonal antibody" refers to a preparation of an antibody molecule having a single molecular composition. Monoclonal antibody compositions exhibit single binding specificity and affinity for a specific epitope.

[0030] The technical solution of the present invention will be described in detail below with reference to specific embodiments. In the following embodiments, unless otherwise specified, the reagents, materials and equipment used can be purchased commercially, prepared by conventional methods, or commonly used in the industry.

[0031] Example 1: Screening and preparation of human anti-RSV F protein monoclonal antibodies 1.1 Labeling of sorted proteins The desired RSV F protein-related specific probe is coupled with a fluorescent dye, as follows: 1.1.1 Recombinant RSV F protein (YK001Ag399 disclosed in CN117886902A) was mixed with biotin at a molecular weight ratio of 1:20. The mixture was incubated at 30°C for 30 min.

[0032] 1.1.2 The reaction product was exchanged with 1× PBS (pH 7.4) using a 30kDa ultrafiltration tube to remove excess biotin.

[0033] 1.1.3 At 20-minute intervals, 1 / 5 molar equivalent of streptavidin-phycoerythrin (SA-PE) was added incrementally to the biotin-labeled antigen-specific probe until the molar ratio of SA-PE to biotin-labeled antigen-specific probe reached 1:1. The probe was incubated at 4°C with gentle shaking. This process yielded fluorescein-labeled probes: Ag(+)-SA-PE. 1.2 Flow cytometry sorting of single antigen-specific B cells PBMCs from recovered patients were separated using the Ficoll-Hypaque density gradient (1.077 g / L) centrifugation method, as follows: step: 1.2.1 Dilute fresh venous blood (heparin anticoagulated) with an equal volume of 1×PBS.

[0034] 1.2.2 Add Ficoll gently to the bottom of the centrifuge tube according to the ratio of peripheral blood to Ficoll of 10:3; spread the diluted blood sample evenly on the surface of the separation medium, keeping the interface between the two liquids clear.

[0035] 1.2.3 At room temperature, centrifuge at 900×g with an acceleration of 4 and a falling speed of 0 for 30 min using a horizontal rotor.

[0036] 1.2.4 After centrifugation, gently aspirate the middle white membrane layer and place it in a new centrifuge tube. Add 10 volumes of 1×PBS, centrifuge at 600×g for 10 min at room temperature, and discard the supernatant. Repeat the washing process once more.

[0037] 1.2.5 Resuspend 1×PBS in complete RPMI 1640 medium for later use and perform viable cell counting.

[0038] Antigen-specific B cells were sorted from PBMCs using flow cytometry. The specific steps are as follows: 1.2.6 Wash cells with 1×PBS and resuspend to a density of 1×10⁻⁶. 6 Add AquaBlue dye at a ratio of 1 μL / mL and incubate at 4°C in the dark for 30 min. Centrifuge at 600 × g for 10 min and discard the supernatant. Repeat the washing process once.

[0039] 1.2.7 PBMCs were stained using flow cytometry with the following antibodies: anti-CD3-PE-Cy7, anti-CD8a-PE-Cy7, anti-CD14-PE-Cy7, anti-CD20-PerCP-Cy5.5, anti-CD19-FITC, anti-IgG-APC-H7, anti-IgM-BV421, anti-CD27-PE-CF594, and Ag(+)-SA-PE, 5 × 10⁻⁶ per molecule. 6 Each PBMC cell was incubated with 2 μL of the above antibody at 4°C in the dark for 40 min, followed by washing 2-3 times with PBS containing 2% BSA. The antibody was then analyzed using BD FACSAria. TM IIICell Sorter flow cytometer for selecting plasma cell-specific cell surface markers (Aqua Blue). - CD3 - CD8a- CD14 - D19 + CD20 + IgG hi IgM lo CD27 + Ag(+) + Ag(-) - Plasma cells were sorted directly into 96-well plates, with each well pre-filled with 20U of RNase inhibitor and 2μL of 1×PBS, and stored at -80℃ for later use. The results of flow cytometry sorting are shown below. Figure 1 , Figure 1 The middle left image shows a delineated population of human blood mononuclear cells; Figure 1 The middle image shows the selection of Aqua Blue from the cells circled in the left image. - CD3 - CD8a - CD14 - D19 + CD20 + IgG hi IgM lo CD27 + cellular; Figure 1 The right-middle figure shows further selection of RSV F cells from the cells circled in the middle figure. + . cells.

[0040] 1.3 Amplification of the variable region gene of the fully human monoclonal antibody using single-cell PCR technology 1.3.1 Reverse Transcription PCR: Referring to the instruction manual (TaKaRa, 6210A), the procedure is briefly described below: Eight single cells were sorted using flow cytometry. To each reaction system, add 4 μL of 5× buffer, 1.1 μL of dNTPs, 1 μL of reverse transcriptase, 1 μL of Oligo dT primer, and water to a final volume of 20 μL. The reverse transcription PCR conditions were: incubation at 65°C for 5 min, followed by rapid cooling on ice. Then, reverse transcription was performed at 42°C for 60 min, followed by pre-denaturation at 95°C for 5 min, and then cooling on ice.

[0041] 1.3.2 Nested PCR Using 4 μL of the reverse transcription product as a template, PCR was performed to amplify the variable regions of H, κ, and λ. The primers for amplifying the variable regions of the heavy chain, kappa light chain, and λ light chain are shown in Table 1 below.

[0042] Table 1. Primers for nested PCR

[0043] The PCR reaction system includes: 25 μL of 2× DNA polymerase buffer, primers as shown above, 2 μL of template (the first round contains the reverse transcription product, and the second round contains the PCR product from the first round), and water to a final volume of 50 μL. The PCR reaction conditions are: 94℃ pre-denaturation for 4 min, followed by 94℃ for 30 s, 58℃ (H, λ strand) or 60℃ (κ strand) for 30 s, 72℃ for 45 min, 50 cycles, and a final extension at 72℃ for 10 min.

[0044] 1.3.3 Agarose gel electrophoresis A clone in which both heavy and light chain genes were successfully amplified in a single cell was considered a successfully paired clone. 5 μL of nested PCR amplification product was subjected to 2% agarose gel electrophoresis. Results are as follows... Figure 2 As shown, lane 1 contains the PCR product of the heavy chain variable region Hc, with an amplified fragment of approximately 450 bp. Paired positive clones were ligated into a T vector and sequenced. The antibody variable region sequences obtained from sequencing were analyzed using Vector NTI software and logged into the IMGT website.

[0045] 1.4 Construction of a eukaryotic expression vector for a fully human monoclonal antibody against RSV F protein 1.4.1 The above light chain / heavy chain genes were inserted into the monoclonal restriction sites of the pcDNA3.1(+ / -) expression vector (purchased from Invitrogen) to construct expression vectors for fully human monoclonal antibodies against RSV F protein.

[0046] 1.4.2 Transform DH5α competent bacteria with the above ligation product and incubate overnight at 37°C on a plate containing ampicillin.

[0047] 1.4.3 Ten single colonies were selected and PCR was performed using specific primers. The reaction conditions were: 94℃ pre-denaturation for 3 min, 94℃ denaturation for 30 s, 55℃ annealing for 30 s, 72℃ extension for 1 min 40 s, 28 cycles, and a final extension at 72℃ for 5 min. 1.4.4 Five μL of the PCR product was electrophoresed on a 1% agarose gel. Transformants containing the antibody heavy or light chain gene were identified among the positive transformants. The results showed that the recombinant expression vector of RSV F protein viral monoclonal antibody heavy / light chain constructed in this invention had the correct sequence.

[0048] 1.5 Transient expression of monoclonal antibodies and affinity chromatography and molecular sieve purification 1.5.1 Using the Expi293 expression system, 15 μg of heavy chain expression plasmid and 15 μg of light chain expression plasmid were mixed and transfected into Expi293F cells according to the manufacturer's instructions (ThermoFisher Scientific, A14635). Cell status was monitored after transfection, and cells were harvested when cell viability dropped to 70%. After centrifugation at 5000 rpm / min for 40 min, the supernatant was collected by filtering through a 0.22 μm bottleneck filter.

[0049] 1.5.2 Affinity chromatography purification was performed using a 15 mL fresh Protein A affinity chromatography column. Before sample loading, the column was equilibrated with 8 column volumes of 1×PBS buffer, and the sample was injected after the conductivity reached baseline. After sample loading, the column was washed with 8 column volumes of 1×PBS buffer until the baseline stabilized. The target protein was eluted fractionally with Elution buffer. Once the OD280 approached baseline, collection was stopped, and 1 mL of 1M Tris (pH 9.0) was added to each eluent. The eluted protein was concentrated by ultrafiltration using an ultrafiltration tube with a molecular weight cutoff of 30 kDa, and its concentration was measured using Nanodrop.

[0050] 1.5.3 The antibody obtained by ultrafiltration was further purified using a Superdex 200 molecular sieve column. The collected peaks of the molecular sieve purification were combined, concentrated using an ultrafiltration tube with a molecular weight cutoff of 30 kDa, and the concentration was detected by Nanodrop.

[0051] 1.5.4 The purified monoclonal antibody was analyzed by SDS-PAGE, and the results are shown in the figure. Figure 4 , Figure 5 , Figure 4 This is the result of antibody purification by gel filtration chromatography. Figure 5 These are the results of SDS-PAGE assays, with lane 1 showing the results of non-reducing SDS-PAGE and lane 2 showing the results of reduced SDS-PAGE. In reduced electrophoresis, the expected molecular weights of the heavy and light chains were 50 kDa and 25 kDa, respectively, with lane M indicating the molecular weight. In non-reducing electrophoresis, the expected molecular weight of the full-molecule monoclonal antibody was 150 kDa, which is consistent with expectations.

[0052] Example 2: Detection of C16's neutralizing ability for RSV using a trace neutralization method 2.1 Antibody dilution: Dilute the antibody to the required concentration using DMEM medium. The initial concentration in each well is 100 μg / mL (100 μL system). Add 25 μL to each well, and dilute at a ratio of 1:3. Set up two replicates for each gradient.

[0053] 2.2 Virus dilution: Prepare the virus using DMEM to the required concentration, mix well, and add 25 μL to each well, starting from the second row of wells. Incubate at 37°C for 1 hour. The first row of wells contains positive and negative controls without virus.

[0054] 2.3 Dilute the cells to 4 × 10⁻⁶ 5 Cells / ml, add 50 μL of cell suspension to each well and mix with antibody-virus mixture. Incubate in a cell culture incubator at 37°C and 5% CO2 for 24 h, then add 50 μL of 10% FBS DMEM and continue incubation for another 24 h. Observe and count the virus neutralization effect under a fluorescence microscope.

[0055] 2.4 Statistical Analysis: Cells without virus and antibodies were used as blank controls, and cells without antibodies were used as virus controls. The neutralization percentage was calculated as (sample signal - blank control signal) / (virus control signal - blank control signal) × 100%. Statistical analysis was performed using GraphPad Prism 7.

[0056] 2.5 Results: Figure 6 The neutralizing effect of different antibody concentrations on pseudoviruses was demonstrated. The C16 monoclonal antibody disclosed in this invention has an IC50 of 153.10 ng / ml in a viral model.

[0057] The sequences involved in the embodiments are shown below: SEQ ID NO.1:GFTLSSYA SEQ ID NO.2: ISFDGSNE SEQ ID NO.3: ARDNPHLESGTYRGDAFDI SEQ ID NO.4:QPITSY SEQ ID NO.5:QQSFLPYT SEQ ID NO.6:QVQLVESGGGVVQPGRSLRLSCAAS SEQ ID NO.7:MHWVRQAPGKGLEWVAV SEQ ID NO.8:YHADSVKGRFTVSRDNSRNTVSLQMTNVGTDDTALYYC SEQ ID NO.9:WGQGTMVTVSS SEQ ID NO.10:MPQSPFSLSSASVGDRVTITCRAS SEQ ID NO.11:LNWYQKKPGKAPKLLIY SEQ ID NO.12:TLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC SEQ ID NO.13:FGQGTNLEIK SEQ ID NO.14:QVQLVESGGGVVQPGRSLRLSCAASGFTLSSYAMHWVRQAPGKGLEWVAVISFDGSNEYHADSVKGRFTVSRDNSRNTVSLQMTNVGTDDTALYYCARDNPHLESGTYRGDAFDIWGQGTMVTVSS SEQ ID NO.15:MPQSPFSLSASVGDRVTITCRASQPITSYLNWYQKKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSFLPYTFGQGTNLEIK SEQ ID NO.16:CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCCTCAGTTCCTATGCTATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATTTGATGGAAGTAATGAATATCACGCTGACTCCGTGAAGGGCCGATTCACCGTCTCCAGAGACAATTCCAGGAACACGGTGTCTCTGCAAATGACCAACGTGGGAACTGACGACACGGCTCTATATTACTGCGCGAGAGATAATCCCCACCTCGAGAGTGGGACCTACCGGGGGGATGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCAG SEQ ID NO.17:ATGCCCCAGTCTCCATTCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGCCCATTACCAGTTATTTAAATTGGTATCAGAAGAAGCCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCACTTT GCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACGGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTTCTTACCGTACACTTTTGGCCAGGGGACCAACCTGGAGATCAAAC The embodiments provided above are not intended to limit the scope of the invention, nor are the described steps intended to limit the order of execution. Any obvious modifications made to the invention by those skilled in the art based on existing common knowledge also fall within the scope of protection defined by the claims.

Claims

1. A fully human neutralizing monoclonal antibody against RSVF protein, characterized in that, The monoclonal antibody is composed of a heavy chain and a light chain. The amino acid sequences of the three CDR regions of the variable region of the heavy chain, namely CDR1, CDR2 and CDR3, are shown in SEQ ID NO. 1, 2 and 3, respectively. The amino acid sequences of the three CDR regions of the variable region of the light chain, namely CDR1, are shown in SEQ ID NO. 4, CDR2, and CDR3, are shown in SEQ ID NO.

5.

2. The monoclonal antibody according to claim 1, characterized in that, In the monoclonal antibody, the amino acid sequences of the four FR regions of the heavy chain: FR1, FR2, FR3, and FR4 are shown in SEQ ID NO. 6, 7, 8, and 9, respectively; and the amino acid sequences of the four FR regions of the light chain: FR1, FR2, FR3, and FR4 are shown in SEQ ID NO. 10, 11, 12, and 13, respectively.

3. The monoclonal antibody according to claim 1 or 2, characterized in that, In the monoclonal antibody, the heavy chain has the amino acid sequence shown in SEQ ID NO.14, and the light chain has the amino acid sequence shown in SEQ ID NO.

15.

4. A nucleic acid molecule encoding the monoclonal antibody of any one of claims 1-3.

5. The nucleic acid molecule according to claim 4, characterized in that, The nucleic acid molecule encoding the heavy chain in the monoclonal antibody has: (a) a nucleotide sequence as shown in SEQ ID NO.16; or, (b) a complementary nucleotide sequence to the nucleotide sequence shown in SEQ ID NO.16; or, a nucleotide sequence that encodes the same protein as (a) or (b) but is different from the nucleotide sequence of (a) or (b) due to the degeneracy of the genetic code.

6. The nucleic acid molecule according to claim 4, characterized in that, The nucleic acid molecule encoding the light chain in the monoclonal antibody has: (c) a nucleotide sequence as shown in SEQ ID NO.17; or, (d) a complementary nucleotide sequence to the nucleotide sequence shown in SEQ ID NO.17; or, a nucleotide sequence that encodes the same protein as the nucleotide sequence in (c) or (d), but is different from the nucleotide sequence in (c) or (d) due to the degeneracy of the genetic code.

7. A vector expressing the monoclonal antibody according to any one of claims 1-3.

8. A cell expressing the monoclonal antibody according to any one of claims 1-3.

9. A drug for treating respiratory syncytial virus (RSV), characterized in that, The drug comprises the monoclonal antibody as described in any one of claims 1-3, and pharmaceutically acceptable excipients.

10. A diagnostic kit for diagnosing respiratory syncytial virus, characterized in that, The kit contains the monoclonal antibody as described in any one of claims 1-3.

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